鹅掌楸LcKNOX3转录因子基因全长克隆及组织表达特异性分析  

Full Length Cloning and Tissue Expression Specific Analysis of LcKNOX3 Transcription Factor Gene in Liriodendron chinense

作　　者：马际凯 成彦丽 仲维平[1] 郝自远 李火根[1] Ma Jikai;Cheng Yanli;Zhong Weiping;Hao Ziyuan;Li Huogen(Co-Innovation Center for Sustainable Forestry in Southern China,Nanjing Forestry University,Nanjing,210037)

机构地区：[1]南京林业大学南方现代林业协同创新中心,南京210037

出　　处：《分子植物育种》2019年第2期437-444,共8页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31770718; 31470660);江苏省高校优势学科(PAPD)共同资助

摘　　要：KNOX基因是一类同源异形盒Homeobox gene转录因子家族基因,在植物的生长发育过程中发挥重要作用并与植物形态建成密切相关。本研究利用RACE技术从鹅掌楸叶芽中克隆出1个KNOX基因的全长序列,并将该基因序列与拟南芥KNOX类基因进行比对。通过生物信息学分析预测该基因的开放阅读框(ORF)以及对基因编码蛋白结构进行分析,与其他物种的同类基因进行同源性比对,构建系统进化树;同时,采用qReal Time-PCR技术对该基因进行鹅掌楸组织表达特异性分析。结果表明,LcKNOX3基因全长1 732 bp,开放阅读框(ORF)长945 bp,编码314个氨基酸,蛋白分子量为34 848.89 kD,等电点(pI)为5.51,不稳定系数为47.52,归类为不稳定蛋白,编码蛋白结构具有4个TALE蛋白家族的保守结构域。BLAST比对结果显示,该基因与拟南芥KNAT3基因相似度最高,将其命名为LcKNOX3。利用Phyre2软件预测该基因编码蛋白的结构,发现与Homeodomain-like蛋白最为相似,系统进化树分析发现与凤梨(Ananas comosus)进化关系较近。根据组织表达特异性分析结果,发现LcKNOX3基因在鹅掌楸的花瓣和茎中的表达量最高,从高到低的顺序为:花瓣>茎>雄蕊>雌蕊>芽>花芽>叶。本研究结果可为鹅掌楸形态建成相关基因的鉴定提供参考依据。KNOX genes belong to homeobox tra nscription factor family which plays a significant role in plant development and relate to plant morphogenesis very closely. In this paper, we used RACE technical method to clone a full length of KNOX gene that be named LcKNOX3 gene, and compared with Arabidopsis KNOX genes. By bioinformatics analysis, we predicted open reading frame(ORF) of LcKNOX3 gene and analyzed the structure of gene coding protein, and then we build phylogenetic trees through homology comparison with other similar gene.What’s more, qReal-Time PCR was used to investigate expression of different issues in Liriodendron chinense. All the results showed that the length of Lc KNOX3 gene was 1 732 bp and ORF was 945 bp, coding 314 amino acids.The protein, containing four conserved domain of TALE family, been sorted to unstable protein, has 34 848.89 kD molecular weight, 5.51 pI and 47.52 coefficient of instability. The results of BLAST compared showed that the gene named LcKNOX3 had the highest similarity. This study predicted the cording protein of LcKNOX3 by Phyre2,and found that this gene has most similar with Homeodomain-like protein. According to phylogenetic trees, it was closed to Ananas comosus in evolutional relationship. The specific tissues expression specific analysis showed that LcKNOX3 genes expression in petals and stem are higher than other tissues, and the expression order from high to low was that petals >stem >stamen >pistil >flower bud >bud >leaf. This research results would provide some basic theories about KNOX genes function and organ morphogenesis in Liriodendron chinense.

关 键 词：LcKNOX3基因 鹅掌楸 生物信息学分析 组织表达 

分 类 号：S792.21[农业科学—林木遗传育种]